LOCUS       sequence-308137-        5868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of MS2 Virus Like Particles packaged with the sequence 
            for the SARS-CoV-2 Nucleocapsid Phosphoprotein gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5868)
  AUTHORS   Crone MA, Priestman M, Ciechonska M, Jensen K, Sharp DJ, Anand A, 
            Randell P, Storch M, Freemont PS
  TITLE     A role for Biofoundries in rapid development and validation of 
            automated SARS-CoV-2 clinical diagnostics.
  JOURNAL   Nat Commun. 2020 Sep 8;11(1):4464. doi: 10.1038/s41467-020-18130-3.
  PUBMED    32900994
REFERENCE   2  (bases 1 to 5868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-09-8"; volume: "11(1):4464. doi"; pages: " 
            10.1038/s41467-020-18130-3"
COMMENT     Sequence Label: pMC059-HisMS2_PLP_NucPhos_pac
FEATURES             Location/Qualifiers
     source          1..5868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1253..1639
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1682..1699
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2078..2125
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2171..2189
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3450..3468
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      3483..3530
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3628..3645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3738..4323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3818..3837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4497..5291)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5292..5396)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5697..5719
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5711..5728
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5712..5728
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5733..5751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5752..5776
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        join(5785..5790,5791..5808,5809..5814)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             5834..5856
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atcgctgtag gtagccggaa ttccattcct aggaggtttg acctgtgcga gcttttagta
       61 cccttgatag ggagaacgaa accttcgtcc cctccgttcg cgtttacgcg gacggtgaga
      121 ctgaagataa ctcattctct ttaaaatatc gttcgaactg gactcccggt cgttttaact
      181 cgactggggc caaaacgaaa cagtggcact acccctctcc gtattcacgg ggggcgttaa
      241 gtgtcacatc gatagatcaa ggtgcctaca agcgaagtgg gtcatcgtgg ggtcgcccgt
      301 acgaggagaa agccggtttc ggcttctccc tcgacgcacg ctcctgctac agcctcttcc
      361 ctgtaagcca aaacttgact tacatcgaag tgccgcagaa cgttgcgaac cgggcgtcga
      421 ccgaagtcct gcaaaaggtc acccagggta attttaacct tggtgttgct ttagcagagg
      481 ccaggtcgac agcctcacaa ctcgcgacgc aaaccattgc gctcgtgaag gcgtacactg
      541 ccgctcgtcg cggtaattgg cgccaggcgc tccgctacct tgccctaaac gaagatcgaa
      601 agtttcgatc aaaacacgtg gccggcagat ggttggagtt gcagttcggt tggttaccac
      661 taatgagtga tatccagggt gcatatgaga tgcttacgaa ggttcacctt caagagtttc
      721 ttcctatgag agccgtacgt caggtcggta ctaacatcaa gttagatggc cgtctgtcgt
      781 atccagctgc aaacttccag acaacgtgca acatatcgcg acgtatcgtg atatggtttt
      841 acataaacga tgcacgtttg gcatggttgt cgtccctagg tatcttgaac ccactaggta
      901 tagtgtggga aaaggtgcct ttctcattcg ttgtcgactg gctcctacct gtaggtaaca
      961 tgctcgaggg ccttacggcc cccgtgggat gctcctacat gtcaggaaca gttactgacg
     1021 taataacggg tgagtccatc ataagcgttg acgctcccta cgggtggact gtggagagac
     1081 agggcactgc taaggcccaa atctcagcca tgcatcgagg ggtacaatcc gtatggccaa
     1141 caactggcgc gtacgtaaag tctcctttct cgatggtcca taccttagat gcgttagcat
     1201 taatcaggca acggctctct agatagagcc ctcaaccgga gtttgaagca tggcttctaa
     1261 ctttactcag ttcgttctcg tcgacaatgg cggaactggc gacgtgactg tcgccccaag
     1321 caacttcgct aacggggtcg ctgaatggat cagctctaac tcgcgttcac aggcttacaa
     1381 agtaacctgt agcgttcgtc agagctctgc gcagaatcgc aaatacacca tcaaagtcga
     1441 ggtgcctaaa gtggcaaccc agactgttgg tggtgtagag cttcctgtag ccgcatggcg
     1501 ttcgtactta aatatggaac taaccattcc aattttcgct acgaattccg actgcgagct
     1561 tattgttaag gcaatgcaag gtcttctaaa agatggaaac ccgattccct cagcaatcgc
     1621 agcaaactcc ggcatctacg ctaactttac tcagttcgtt ctcgtcgaca atggcggtac
     1681 ccatcaccat caccatcacg gtaccggcga cgtgactgtc gccccaagca acttcgctaa
     1741 cggggtcgct gaatggatca gctctaactc gcgttcacag gcttacaaag taacctgtag
     1801 cgttcgtcag agctctgcgc agaatcgcaa atacaccatc aaagtcgagg tgcctaaagt
     1861 ggcaacccag actgttggtg gtgtagagct tcctgtagcc gcatggcgtt cgtacttaaa
     1921 tatggaacta accattccaa ttttcgctac gaattccgac tgcgagctta ttgttaaggc
     1981 aatgcaaggt cttctaaaag atggaaaccc gattccctca gcaatcgcag caaactccgg
     2041 catctactaa tagacgccgg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2101 taaacgggtc ttgaggggtt ttttgtcgaa cagaaagtaa tcgtattgta cacggccgca
     2161 taatcgaaat taatacgact cactatagga tgtctgataa tggaccccaa aatcagcgaa
     2221 atgcaccccg cattacgttt ggtggaccct cagattcaac tggcagtaac cagaatggag
     2281 aacgcagtgg ggcgcgatca aaacaacgtc ggccccaagg tttacccaat aatactgcgt
     2341 cttggttcac cgctctcact caacatggca aggaagacct taaattccct cgaggacaag
     2401 gcgttccaat taacaccaat agcagtccag atgaccaaat tggctactac cgaagagcta
     2461 ccagacgaat tcgtggtggt gacggtaaaa tgaaagatct cagtccaaga tggtatttct
     2521 actacctagg aactgggcca gaagctggac ttccctatgg tgctaacaaa gacggcatca
     2581 tatgggttgc aactgaggga gccttgaata caccaaaaga tcacattggc acccgcaatc
     2641 ctgctaacaa tgctgcaatc gtgctacaac ttcctcaagg aacaacattg ccaaaaggct
     2701 tctacgcaga agggagcaga ggcggcagtc aagcctcttc tcgttcctca tcacgtagtc
     2761 gcaacagttc aagaaattca actccaggca gcagtagggg aacttctcct gctagaatgg
     2821 ctggcaatgg cggtgatgct gctcttgctt tgctgctgct tgacagattg aaccagcttg
     2881 agagcaaaat gtctggtaaa ggccaacaac aacaaggcca aactgtcact aagaaatctg
     2941 ctgctgaggc ttctaagaag cctcggcaaa aacgtactgc cactaaagca tacaatgtaa
     3001 cacaagcttt cggcagacgt ggtccagaac aaacccaagg aaattttggg gaccaggaac
     3061 taatcagaca aggaactgat tacaaacatt ggccgcaaat tgcacaattt gcccccagcg
     3121 cttcagcgtt cttcggaatg tcgcgcattg gcatggaagt cacaccttcg ggaacgtggt
     3181 tgacctacac aggtgccatc aaattggatg acaaagatcc aaatttcaaa gatcaagtca
     3241 ttttgctgaa taagcatatt gacgcataca aaacattccc accaacagag cctaaaaagg
     3301 acaaaaagaa gaaggctgat gaaactcaag ccttaccgca gagacagaag aaacagcaaa
     3361 ctgtgactct tcttcctgct gcagatttgg atgatttctc caaacaattg caacaatcca
     3421 tgagcagtgc tgactcaact caggcctaaa catgaggatc acccatgtcc gctgagcaat
     3481 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg taatttcact
     3541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taacatggtc atagctgttt
     3601 ccttgcgtat tgggcgctct ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3661 ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgc tacagagttc
     4141 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttattag
     4501 aaaaattcat ccagcagacg ataaaacgca atacgctggc tatccggtgc cgcaatgcca
     4561 tacagcacca gaaaacgatc cgcccattcg ccgcccagtt cttccgcaat atcacgggtg
     4621 gccagcgcaa tatcctgata acgatccgcc acgcccagac ggccgcaatc aataaagccg
     4681 ctaaaacggc cattttccac cataatgttc ggcaggcacg catcaccatg ggtcaccacc
     4741 agatcttcgc catccggcat gctcgctttc agacgcgcaa acagctctgc cggtgccagg
     4801 ccctgatgtt cttcatccag atcatcctga tccaccaggc ccgcttccat acgggtacgc
     4861 gcacgttcaa tacgatgttt cgcctgatga tcaaacggac aggtcgccgg gtccagggta
     4921 tgcagacgac gcatggcatc cgccataatg ctcacttttt ctgccggcgc cagatggcta
     4981 gacagcagat cctgacccgg cacttcgccc agcagcagcc aatcacggcc cgcttcggtc
     5041 accacatcca gcaccgccgc acacggaaca ccggtggtgg ccagccagct cagacgcgcc
     5101 gcttcatcct gcagctcgtt cagcgcaccg ctcagatcgg ttttcacaaa cagcaccgga
     5161 cgaccctgcg cgctcagacg aaacaccgcc gcatcagagc agccaatggt ctgctgcgcc
     5221 caatcatagc caaacagacg ttccacccac gctgccgggc tacccgcatg caggccatcc
     5281 tgttcaatca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5401 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5461 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5521 aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc ccattcgcca
     5581 ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca
     5641 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5701 gtcacgacgt tgtaaaacga cggccagtga ggtaatacga ctcactatag gggaattgtg
     5761 agcggataac aattcccctc tagatttaca ctttatgctt ccggctcgta tgtttctaga
     5821 gaattcaaat aattttgttt aactttaaga aggagattta aattggct
//
